Heavy Metals in the Muscle and Hepatopancreas of Red Swamp Crayfish (Procambarus clarkii) in Campania (Italy).

The aim of this study was to carry out a quali-quantitative analysis of the presence of non-essential and essential trace elements in freshwater crayfish (Procambarus clarkii) edible tissues to establish the healthiness of this product and to evaluate the pollution status of the sampling sites included in the present study. P. clarkii is one of the most common species of freshwater crustaceans in Italy, regularly consumed by local people. Moreover, the crayfish, due to its trophic position and diet, can be considered as an excellent bioindicator of the health status of the ecosystem. We collected sixty crayfish samples from two different sites in Campania (Italy): Villa Literno and Sessa Aurunca. Concentrations of trace elements were determined by Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). Our data showed low concentrations of Cd, Hg and Pb, with values below the European Commission MRL (Commission Regulation (EC) 1881/2006). We suggest that data obtained from this study showed that crayfish collected from Villa Literno and Sessa Aurunca were safe for human consumption. Furthermore, the results of this research indicated mild contamination of heavy metals of the sampling sites, indicating a good health status of the area's aquatic ecosystem.

Diterpenoids from Zhumeria majdae roots as potential heat shock protein 90 (HSP90) modulators.

Four undescribed and 17 known diterpenoids were isolated from the roots of Zhumeria majdae Rech.f. & Wendelbo. Using 1D and 2D NMR spectroscopy, ECD spectroscopy, and HRESIMS data analysis, the structures of the undescribed compounds were elucidated. The anti-proliferative activity of isolated compounds was evaluated against HeLa and MCF7 cancer cell lines. The binding affinity of all compounds to HSP90, one of the targets for the modern anticancer therapy, was investigated using surface plasmon resonance. The results demonstrated that lanugon Q interacted with the chaperone. To explain its mechanism of action, experimental and computational tests were also conducted.

Systematic Phytochemical Screening of Different Organs of Calotropis procera and the Ovicidal Effect of Their Extracts to the Foodstuff Pest Cadra cautella.

In developing countries, crop deterioration is mainly caused by inappropriate storage conditions that promote insect infestation. Synthetic pesticides are associated with serious adverse effects on humans and the environment. Thus, finding alternative "green" insecticides is a very pressing need. Calotropis procera (Aiton) Dryand (Apocynaceae) growing in Saudi Arabia was selected for this purpose. LC-MS/MS analysis was applied to investigate the metabolic composition of different C. procera extracts. Particularly, C. procera latex and leaves showed a high presence of cardenolides including calactin, uscharidin, 15ß-hydroxy-calactin, 16ß-hydroxy-calactin, and 12ß-hydroxy-calactin. The ovicidal activity of the extracts from different plant organs (flowers, leaves, branches, roots), and of the latex, against Cadra cautella (Walker) (Lepidoptera, Pyralidae) was assessed. Extracts of C. procera roots displayed the most potent activity with 50% of C. cautella eggs not hatching at 10.000 ppm (1%).

Antiprotozoal activity of diterpenoids isolated from Zhumeria majdae- absolute configuration by circular dichroism.

PURPOSE: Zhumeria majdae, a unique species of the Zhumeria genus, is an endemic Iranian plant in the Lamiaceae family. Phytochemical investigation and biological activity of this plant are rarely reported. The current study aimed to find new antiprotozoal compounds from the roots of Z. majdae and to determine the absolute configuration of isolated compounds by circular dichroism. METHODS: The extraction process from roots and aerial parts of Z. majdae was carried out by hexane, ethyl acetate and methanol followed by testing their antiprotozoal effects against Leishmania donovani, Trypanosoma brucei rhodesiense, T. cruzi, and Plasmodium falciparum, respectively. Structure elucidation was done using 1D and 2D NMR spectroscopy and HREIMS spectrometry. In addition, experimental and theoretical circular dichroism spectroscopy was used to establish absolute configuration. RESULTS: In comparison with aerial parts, the hexane extract from roots showed superior activity against T. b. rhodesiense, L. donovani and P. falciparum with IC50 values of 5.4, 1.6 and 2.1 µg/ml, respectively. From eight abietane-type diterpenoids identified in roots, six were reported for the first time in the genus Zhumeria. 11,14-dihydroxy-8,11,13-abietatrien-7-one (6) exhibited a promising biological activity against P. falciparum (IC50 8.65 µM), with a selectivity index (SI) of 4.6, and lanugon Q (8) showed an IC50 value of 0.13 µM and SI of 15.4 against T. b. rhodesiense. CONCLUSION: Altogether, according to the results, of 8 isolated compounds, dihydroxy-8,11,13-abietatrien-7-one (6) and lanugon Q (8) exhibited a promising activity against T. b. rhodesiense and P. falciparum. In conclusion, these compounds could be potential candidates for further analysis and may serve as lead compounds for the synthesis of antiprotozoal agents. Graphical abstract.

Fusicoccane Diterpenes from Hypoestes forsskaolii as Heat Shock Protein 90 (Hsp90) Modulators.

Ten new (1-10) and six known (11-16) fusicoccane diterpenes were isolated from the roots of Hypoestes forsskaolii. The structural characterization of 1-10 was performed by spectroscopic analysis, including 1D and 2D NMR, ECD, and HRESIMS experiments. From a perspective of obtaining potential Hsp90α inhibitors, the isolates were screened by surface plasmon resonance measurements and their cytotoxic activity was assayed using Jurkat and HeLa cancer cells. Compound 6, 18-hydroxyhypoestenone, was shown to be the most active compound against Hsp90, and its interactions were studied also by biochemical and cellular assays and by molecular docking.

Spartium junceum L. poisoning in small ruminants.

An outbreak of neurological disorders in a flock of 20 sheep coming from a rural farm in Civitella Roveto, Italy, occurred in winter 2015. All the animals showed tonic-clonic convulsions followed by muscle paralysis associated with dilated pupils, tremor, tachycardia, tachypnea and diarrhea. The presence of bundles of dry broom of Spartium junceum L. in the feed, eaten by the animals supported the hypothesis of plant intoxication. Two animals died after worsening of clinical signs. The anatomopathological findings and the laboratory results ruled out viral or bacterial infections or accidental exposure to other toxics. Phytochemical study showed the presence of large amount of cytisine, a nicotinic acetylcholine receptor agonist, in all parts of the plant eaten by the animals. Clinical and pathological findings, the complete remission of clinical signs after the exclusion of dry broom from the diet, together with the results of phytochemical analyses results corroborated the hypothesis of S. junceum L. intoxication.

Antioxidant Potential of Herbal Preparations and Components from Galactites elegans (All.) Nyman ex Soldano.

Galactites is a genus of flowering plants belonging to Asteraceae family. This genus is mainly represented by the Galactites elegans (All.) Nyman ex Soldano, the milky thistle, a plant of Mediterranean origin. Galactites elegans is consumed as a monofloral boar thistle honey. Chromatography separation of CHCl3 and n-BuOH extracts of aerial parts of G. elegans led to isolation of 18 pure compounds. Their structures were elucidated by 1D-and 2D-NMR spectroscopy and confirmed by mass spectrometry analysis. Sinapic aldehyde, abietin, chlorogenic acid, neochlorogenic acid, 8α-hydroxypinoresinol, 9α-hydroxypinoresinol, pinoresinol, 4-ketopinoresinol, nortrachelogenin, and erythro-guaiacylglycerol-ß-O-4'-dihydroconiferyl alcohol were isolated from CHCl3 extract, while luteolin 4'-O-glucuronide, naringenin-7-O-neohesperidoside, kaempferol-3-O-α-L-rhamnopyranosyl-(1→6)-ß-D-glucopyranoside, apigenin-7-O-α-L-rhamnopyranosyl-(1→6)-ß-D-glucopyranoside, quercitrin, quercetin-3-O-α-L-rhamnopyranosyl-(1→6)-ß-D-glucopyranoside, ciwujiatone, and nortrachelogenin-4,4'-di-O-ß-D-glucopyranoside were obtained from n-BuOH extract. The majority of isolated compounds displayed a significant antioxidant potential in vitro test (DPPH). The ability of compounds to reduce the level of peroxides in control and BHP-treated Jurkat cells was studied. The lignan derivatives were also able to reduce at 50 µM the basal level of peroxides in Jurkat cells as well as counteract peroxide increase induced by BHP treatment. Particularly 8α-hydroxypinoresinol was the most active showing 70% of peroxide level inhibition.

In Vitro Anthelminthic Efficacy of Hypoestes forskaolii (Vahl) R.Br (Acanthaceae) Extracts on Gastrointestinal Nematodes of Sheep.

The anthelminthic efficacy of the crude extracts of Hypoestes forskaolii (Vahl) R.Br (Acanthaceae) against gastrointestinal nematodes (GIN) in sheep was investigated using the in vitro egg hatch inhibition assay. Faecal samples were collected from sheep with naturally occurring infection of GIN (Trichostrongylus spp., Chabertia ovina, Cooperia spp., Haemonchus contortus and Teladorsagia spp.). Crude leaf extracts of H. forskaolii was obtained using increasing polarity solvents: n-hexane, chloroform, chloroform:methanol 9:1, methanol. Thiabendazole (0.2 µg/mL and 0.5 µg/mL) was used as a positive control and untreated GIN eggs in deionised water served as the negative control. All the extracts exhibited a weak ovicidal activity against GIN (less than 50% of egg hatch). Noteworthy, the n-hexane extract showed a percentage of inhibition of egg hatching greater than other extracts inhibiting the 30.8% at the concentration of 1 mg/mL showing a dose-dependent effect on nematode eggs hatching. Further studies are needed to investigate the effects of extracts used and to evaluate the ovicidal effects of other extracts of H. forskaolii.

Phytochemistry of compounds isolated from the leaf-surface extract of Psiadia punctulata (DC.) Vatke growing in Saudi Arabia.

The surface extract of an accession of Psiadia punctulata (DC.) Vatke (Asteraceae) growing in Saudi Arabia was investigated for its phytochemical composition. A bio-guided investigation of the extract led to the isolation of thirteen ent-kaurane and trachylobane diterpenes and seventeen compounds previously described, including nine flavonoids and eight diterpenes. Three flavonoids and one ent-kaurane diterpene showed antimicrobial activity with MIC100 values ranging from 25 to 150 µg/ml. The extract showed antibacterial activity against Staphylococcus aureus (MIC100 = 180 µg/ml) and antifungal activity against Candida albicans (MIC0 = 130 µg/ml). The isolated 3',4',5,7-tetramethoxyflavone, at a concentration of 40 µg/ml, displayed the ability to reduce biofilm formation of S. aureus and C. albicans by 50% and 90% respectively.

A new glucosidic iridoid from Isodon rubescens

ABSTRACT One new glucosidic iridoid, 6-O-veratroylbarlerin, was isolated from the chloroform/methanol extract of Isodon rubescens (Hemsl.) H.Hara, Lamiaceae aerial parts, along with the known compounds apigenin and caffeic acid. The structure of the new compound was elucidated on the basis of 1D and 2D NMR experiments and ESI-MS technique.

Toxicity of Crepis lacera in grazing ruminants.

BACKGROUND: Crepis lacera is a plant from the Asteraceae family that is common in the Mediterranean region. Farmers believe that this plant may be deadly to small ruminants in areas of southern Italy. However, scientific evidence is lacking, and no proof exists that C. lacera is toxic to ruminants. Necropsies conducted on four sheep revealed lesions in their livers and kidneys. RESULTS: In the current study, we described sheep poisoning and isolated secondary metabolites from Crepis lacera to assess the metabolites' biological activity both in vitro and in vivo. Phytochemical study of the aerial portions of Crepis lacera led to the isolation of five sesquiterpene lactones and two phenolic compounds. Cellular viability was evaluated in cell cultures of the bovine kidney cell line Madin Darby Bovine Kidney (MDBK) after incubation with phytochemicals. Our results showed that three sesquiterpene lactones, 8-epidesacylcynaropicrin-3-O-ß-glucopyranoside (2), 8-epigrosheimin (3), and 8-ß-hydroxydehydrozaluzanin C (4), were cytotoxic after 48 h of incubation. In addition, in the in vivo study, animals that received 1 mg/kg body weight (bw) of Crepis lacera extract and were then sacrificed after 48 h showed significant lesions in their liver, lungs and kidneys. These lesions were also found in rats that received 2 mg/kg bw of the same extract and sacrificed after 24 and 48 h. CONCLUSIONS: These results validate the hypothesis that C. lacera is potentially dangerous when ingested in large quantities by grazing small domestic ruminants. Further studies are necessary to clarify the molecular mechanisms of Crepis spp. toxicity in animals.

A Novel Tirucallane-type Triterpene and Sesquiterpene from Trichilia maynasiana.

One new tirucallane-type triterpene {3ß, 24-dihydroxytirucallan-7,25-diene, 24-sulfate (1)}, one new sesquiterpene {7-epi-10-hydroxychabrol-1(2)-en-4,5-dione A (2)}, together with three known tirucallanes, and four aromadendranes were isolated from the leaves of Trichilia maynasiana C. DC.. Their structures were determined by means of NMR spectroscopy, mass spectrometric analysis, and chemical methods.

Sesquiterpene Lactones from Vernonia nigritiana.

Two new sesquiterpenes, 8α-(4-hydroxymethacryloyl)-14-acetoxy-salonitenolide (1) and 8α-(2-hydroxymethyl 2-butenoyl)-14-acetoxy-salonitenolide (2), together with five known sesquiterpenes were isolated from the leaves of Vernonia nigritiana Oliv. & Hiern. Their structural characterization was obtained on the basis of extensive NMR spectroscopic and mass spectrometric studies.

Phenolic compounds from Limonium pruinosum.

A new phenolic compound, 6'-O-sulfonyl-salidroside (1), together with fifteen known compounds, were isolated from the all parts of Limonium pruinosum. Their structures were elucidated by 1D and 2D NMR experiments including 1D TOCSY, DQF-COSY, HSQC and HMBC spectroscopy, as well as ESIMS analysis. The antioxidant activity of the extracts and all isolated compounds was evaluated by DPPH test.